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      Comparative Evaluation of Different Test Combinations for Diagnosis of Mycobacterium avium Subspecies paratuberculosis Infecting Dairy Herds in India

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          Abstract

          A total of 355 cows were sampled (serum, n = 315; faeces, n = 355; milk, n = 209) from dairy farms located in the Punjab state of India. Faeces and serum/milk samples were screened by acid fast staining and “indigenous ELISA,” respectively. IS 900 PCR was used to screen faeces and milk samples. Bio-load of MAP in dairy cows was 36.9, 15.6, 16.3, and 14.4%, using microscopy, serum ELISA, milk ELISA and milk PCR, respectively. Estimated kappa values between different test combinations: serum and milk ELISA, faecal microscopy and faecal PCR, milk ELISA and milk PCR, faecal PCR and serum ELISA were 0.325, 0.241, 0.682, and 0.677, respectively. Estimation of the relative sensitivity and specificity of different tests in the present study indicated that “serum ELISA” and “milk ELISA” were good screening tests, add “milk PCR” was “confirmatory test” for MAP infection. Combination of milk ELISA with milk PCR may be adopted as a model strategy for screening and diagnosis of JD in lactating/dairy cattle herds in Indian conditions.

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          MEGA3: Integrated software for Molecular Evolutionary Genetics Analysis and sequence alignment.

          S. KUMAR (2004)
          With its theoretical basis firmly established in molecular evolutionary and population genetics, the comparative DNA and protein sequence analysis plays a central role in reconstructing the evolutionary histories of species and multigene families, estimating rates of molecular evolution, and inferring the nature and extent of selective forces shaping the evolution of genes and genomes. The scope of these investigations has now expanded greatly owing to the development of high-throughput sequencing techniques and novel statistical and computational methods. These methods require easy-to-use computer programs. One such effort has been to produce Molecular Evolutionary Genetics Analysis (MEGA) software, with its focus on facilitating the exploration and analysis of the DNA and protein sequence variation from an evolutionary perspective. Currently in its third major release, MEGA3 contains facilities for automatic and manual sequence alignment, web-based mining of databases, inference of the phylogenetic trees, estimation of evolutionary distances and testing evolutionary hypotheses. This paper provides an overview of the statistical methods, computational tools, and visual exploration modules for data input and the results obtainable in MEGA.
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            Practical statistics for medical researched

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              Preclinical and clinical manifestations of paratuberculosis (including pathology).

              The term Johne's disease immediately brings to mind an image of an emaciated, debilitated ruminant with bottlejaw and fluid, pipestream diarrhea. However, the clinical case is merely the tip of the iceberg in terms of the total number of infected animals on the farm. If the clinically affected animal was born on the farm, a minimum of 25 other animals are probably infected and less than 30% of those will be detectable by currently available tests. Infected animals in the early stages of the disease progress slowly over a period of months or several years to stages that may be detected by antigen detection tests and antibody-based diagnostic tests. The duration of each stage of infection depends on age at the time of exposure and the dose of organisms ingested. The lack of tests sensitive enough to detect infected animals in the earliest stages of disease is a serious obstacle to eradication of Johne's disease. The animals with overt infection may be culled from the herd immediately on detection. However, the animals that are shedding organisms below the threshold of detectability and have not mounted an immune response remain behind to contaminate the environment and infect herdmates.
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                Author and article information

                Journal
                Biomed Res Int
                Biomed Res Int
                BMRI
                BioMed Research International
                Hindawi Publishing Corporation
                2314-6133
                2314-6141
                2015
                5 April 2015
                : 2015
                : 983978
                Affiliations
                1Animal Disease Research Centre, Guru Angad Dev Veterinary and Animal Sciences University (GADVASU), Ludhiana, Punjab 141004, India
                2Aquatic Animal Health Division, Central Institute of Brackishwater Aquaculture (CIBA), 75 Santhome High Road, R.A. Puram, Chennai 600028, India
                3Microbiology Laboratory, Animal Health Division, Central Institute for Research on Goats, Makhdoom, P.O. Farah, Mathura, Uttar Pradesh 281122, India
                4Department of Veterinary Medicine, Guru Angad Dev Veterinary and Animal Sciences University (GADVASU), Ludhiana, Punjab 141004, India
                5Division of Animal Biotechnology, Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh 243122, India
                6National JALMA Institute for Leprosy and Other Mycobacterial Diseases (NJIL & OMD), Tajganj, Agra, Uttar Pradesh 282001, India
                7Department of Microbiology, King George's Medical University, Lucknow, Uttar Pradesh 226003, India
                8Amity Institute of Microbial Technology, Amity University Rajasthan, Kant Kalwar, NH-11C Delhi-Jaipur Highway, Jaipur 303 002, India
                9Department of Veterinary Microbiology and Immunology, Uttar Pradesh Pandit Deen Dayal Upadhayay Pashu Chikitsa Vigyan Vishwa Vidyalaya Evam Go-Anusandhan Sansthan (DUVASU), Mathura, Uttar Pradesh 281001, India
                10Division of Pathology, Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh 243122, India
                Author notes
                *Jagdip Singh Sohal: sohal_lucky@ 123456ymail.com

                Academic Editor: Klaus Wimmers

                Author information
                http://orcid.org/0000-0001-9619-7597
                http://orcid.org/0000-0001-7763-5547
                http://orcid.org/0000-0001-7469-4752
                http://orcid.org/0000-0002-2739-8536
                Article
                10.1155/2015/983978
                4402184
                25945351
                8bf7cb4b-82aa-47e8-88f7-49c40a1c34c1
                Copyright © 2015 Rajni Garg et al.

                This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 25 November 2014
                : 24 February 2015
                : 9 March 2015
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                Research Article

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